Utilizing Multielectrode Arrays to Measure Synaptic Physiology in 3D Coculture Spheres

Abstract

Source: Cvetkovic, C., et al. Synaptic Microcircuit Modeling with 3D Cocultures of Astrocytes and Neurons from Human Pluripotent Stem Cells. J. Vis. Exp. (2018)

This video demonstrates the procedure of using the multielectrode array (MEA) to measure synaptic physiology in 3D coculture spheres. This method allows to understand the neural network dynamics and synaptic network burst activity within the 3D spheres.

Protocol

Measurement of Live Synaptic Physiology with Multielectrode Arrays (MEAs)

1. Prepare MEAs for cell culture.

1. Clean the surface of each MEA with 1 mL of detergent for 1 h. Rinse with sterile deionized (DI) water, rinse with 70% ethanol to sterilize, and then air dry in a biosafety cabinet under UV light.
   NOTE: MEAs can be stored in DI water at 4 °C under sterile conditions until use.
2. Prepare a 0.5 mg/mL solution of poly-ornithine (PLO) by dissolving 50 mg of PLO into 100 mL of boric acid buffer. Coat the surface of each MEA with 1 mL of PLO to render it hydrophilic and incuba....

Results

Figure 1: Representative live synaptic physiology of neural spheres on multielectrode arrays (MEAs). (A, A') MEAs are utilized to measure the live synaptic physiology of neural spheres. Spheres of iNeurons (A) or cocultures of iNeuron.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
DMEM/F12	Thermofisher	10565-042	With GlutaMAX supplement
Anti/anti	Thermofisher	15240062
B27	Thermofisher	17504044	Media Supplement
Heparin	Sigma	H3149-50KU	Working concentration: 2 mg/mL
Matrigel membrane matrix	Corning	354230	ECM coating solution. Working concentration: 80 ug/ml. Prepare on ice and ensure that pipettes, tubes, and media are pre-chilled.
MEA 2100 System	Multichannel Systems	MEA2100
N2	Thermofisher	17502048	Media Supplement
Poly-l-ornithine (PLO)	Sigma	P3655-100MG	Working concentration: 0.5 mg/mL